Western Blot Analysis of Liver Proteins

Western blot analysis for Bax, Bcl-2, and AMPK was done as the same method of Park et al. (2020) (link). After homogenizing liver tissues on ice, the tissues were lysed using lysis buffer. Protein content was calculated using Bio-Rad colorimetric protein assay kit (Bio-Rad, Hercules, CA, USA). After separation of 40-μg protein on sodium dodecyl sulfate-poly-acrylamide gels, it was transferred to the nitrocellulose membrane. Then, the reaction mixture was treated with primary antibodies such as mouse β-actin antibody (1:3,000; Santa Cruz Biotechnology), mouse Bcl-2 antibody (1: 1,000; Santa Cruz Biotechnology), rabbit Bax antibody (1:1,000; Santa Cruz Biotechnology), and rabbit AMPK antibody (1:1,000; Santa Cruz Biotechnology). Horseradish peroxidase-conjugated anti-mouse antibodies for β-actin and Bcl-2 and horseradish peroxidase-conjugated anti-rabbit antibodies for Bax and AMPK were used as the secondary antibodies. The expression for bands was analyzed using enhanced chemiluminescence detection system (Santa Cruz Biotechnology).

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Kim S.H., Ko I.G., Jin J.J., Hwang L., Yoon H.S, & Baek S.S. (2022). Treadmill exercise ameliorates ethanol with lipopolysaccharide and carbon tetrachloride-mediated liver injury in mice. Journal of Exercise Rehabilitation, 18(1), 28-33.

Publication 2022

Bio-Rad colorimetric protein assay kit mouse β-actin antibody mouse Bcl-2 antibody rabbit Bax antibody enhanced chemiluminescence detection system

Actin Anti antibodies Antibodies Antibody Assay Bcl 2 Buffer Chemiluminescence Colorimetric Horseradish peroxidase Liver Membrane Mouse Nitrocellulose Poly acrylamide gels Protein Rabbit Sodium dodecyl sulfate Tissues Western blot analysis

Corresponding Organization :

Other organizations : Sangmyung University, Kyung Hee University, Eulji University

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Expression levels of Bax, Bcl-2, and AMPK proteins

control variables

Amount of protein loaded (40 μg)
Antibodies used for Western blot analysis (mouse β-actin, mouse Bcl-2, rabbit Bax, and rabbit AMPK)
Horseradish peroxidase-conjugated secondary antibodies (anti-mouse for β-actin and Bcl-2, anti-rabbit for Bax and AMPK)
Enhanced chemiluminescence detection system used for analysis

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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